微重力培养的新鲜和冻存胰岛联合移植治疗1型糖尿病

目的经微重力培养的新鲜和冻存胰岛联合移植提高1型糖尿病的治疗效果。方法将分离纯化的大鼠胰岛分为（1）体外实验组：实验组1．1：冻存的胰岛经微重力培养；实验组2．1：冻存的胰岛在普通培养基中培养；对照组1：新鲜大鼠胰岛经微重力培养。观察胰岛收获率和体外胰岛素分泌情况。（2）胰岛移植组：即将新鲜和冻存的胰岛经微重力或普通培养7d后分别移植入受体鼠体内，观察移植效果。结果经微重力培养的各组胰岛收获率、DNA含量和胰岛素含量高于普通组。普通培养组在培养后期胰岛素分泌明显下降，并且胰岛素刺激指数明显低于经微重力培养组（P〈0．05）。移植经过微重力培养的500 IEQ新鲜胰岛和1500 IEQ冻存胰岛在移植后1周内可达100％纠正糖尿病，全部受体维持正常血糖耐受曲线一直到观察结束。结论采用细胞内低温保存液（HTS）结合细胞冻存液（DMSO）对大鼠胰岛进行冻存前后经微重力培养可以明显提高胰岛冻存质量，是目前胰岛冻存的最佳选择。使经微重力培养的新鲜和冻存胰岛联合移植一次治愈糖尿病成为可能，并有效的节约了胰岛资源，提高了移植效果。

Treatment of type I diabetes mellitus with co-transplantation of fresh and frozen-thawed islets cultured under microgravity

Objective To investigate the therapeutic effects of co-transplantation of fresh and frozen-thawed islets cultured under microgravity on type I diabetes mellitus.Methods The isolated and purified islets were randomized into: 1) In vitro experimental groups: experimental group 1.1: the frozen-thawed islets were cultured under microgravity;experimental group 2.1: the frozen-thawed islets were cultured with common medium;control group: the fresh islets were cultured under microgravity.The islet recovery and insulin secretion were observed in vitro.2) Transplantation group: After being cultured for 7 d under microgravity or in common medium,the fresh and frozen-thawed islets were implanted into the rat recipients.Then the transplantation effect was observed.Results The islets cultured under microgravity had higher islet recovery rate,DNA level and insulin content than those cultured in common medium.After the culture for 14 d,insulin secreting level significantly declined in common culture groups and the stimulation indices were markedly lower than in the microgravity groups(P<0.05).Co-transplantation of 500 IEQ fresh islets and 1500 IEQ frozen-thawed islets restored blood glucose to normal in all the recipients within 1 week.All the recipients maintained the normal glucose-tolerance curve throughout the observation period.Conclusions Co-cryopreservation of rat islets with HTS and DMSO via culture under microgravity would improve the effect of islet cryopreservation and it is a best choice,which makes it possible to cure diabetes by co-transplantation of fresh and frozen-thawed islets.This method not only greatly saves the islet sources but also improves the transplantation effect.